1. Technical Field
The present invention is related to the construction of an expression vector for the synthesis of a recombinant enzyme. More particularly, the present invention is related to the large scale production of glucocerebrosidase by infecting invertebrate cells with a recombinant baculovirus containing the complete cDNA sequence for encoding glucocerebrosidase.
2. State of the Art
Mutation or deficiency of the lysosomal glycoprotein glucocerebrosidase (EC 3.2.1.45, .beta.-D-glucosyl-N-acylsphingosine glycohydrolase) results in Gaucher's disease. It is estimated that there are about 20,000 cases of this genetic disease in the U.S. alone.
Published methods for producing large quantities of the active human enzyme involve purification of the protein from large amounts of human tissue, such as placenta. It should be noted, however, that the placental glucocerebrosidase has carbohydrate structure different than that of the enzyme found in human liver, spleen, brain or macrophages.
Construction of a cDNA clone containing the entire human glucocerebrosidase coding region has been known (Sorge et al, Proc. Natl. Acad. Sci. USA 82:7289-7293, 1985). However, as it will become clear vide infra, both the cDNA clone of the present invention and the enzyme synthesized therefrom, are qualitatively different from the similar prior art entities.